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Related Concept Videos

LTR Retrotransposons03:08

LTR Retrotransposons

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LTR retrotransposons are class I transposable elements with long terminal repeats flanking an internal coding region. These elements are less abundant in mammals compared to other class I transposable elements. About 8 percent of human genomic DNA comprises LTR retrotransposons. Some of the common examples of LTR retrotransposons are Ty elements in yeast and Copia elements in Drosophila.
The internal coding region of LTR retrotransposons and their mechanism of transposition closely resembles a...
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DNA-only Transposons

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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
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Overview of Transposition and Recombination02:13

Overview of Transposition and Recombination

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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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Non-LTR Retrotransposons03:18

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As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Retroviruses02:33

Retroviruses

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Retroviruses and retrotransposons both insert copies of their genetic elements into the genome of the host cell. Thus, the viral genes are passed on when the host genome is replicated or translated. A typical retroviral DNA sequence contains 3-4 genes that encode the different proteins required for its structural assembly and function as a molecular parasite. This DNA is transcribed into a single mRNA, which is very similar in structure to conventional mRNAs, i.e., it is capped at the 5’...
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Structure and biochemistry-guided engineering of an all-RNA system for DNA insertion with R2 retrotransposons.

KeHuan K Edmonds1,2,3,4,5,6,7, Max E Wilkinson1,2,3,4,5,6,7, Daniel Strebinger1,2,3,4,5,6,7

  • 1Howard Hughes Medical Institute, Cambridge, MA, USA.

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Researchers engineered an all-RNA system for transgene insertion using R2 retrotransposons. This compact system achieves high integration efficiency in human cells, advancing genome engineering tools.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Non-long terminal repeat (non-LTR) retrotransposons, like R2 elements, show promise for transgene insertion.
  • Current applications are hindered by incomplete understanding of retrotransposon mechanisms.

Purpose of the Study:

  • To structurally and biochemically characterize the R2 element from Taeniopygia guttata (R2Tg).
  • To engineer and optimize an all-RNA system for efficient transgene insertion using R2 elements.

Main Methods:

  • Structural and biochemical characterization of R2Tg.
  • Engineering of a donor RNA system by removing non-essential R2 sequences.
  • Chemical modification of donor RNA and delivery optimization.

Main Results:

  • R2Tg cleaves both DNA strands of its ribosomal DNA target.
  • R2Tg utilizes a pseudoknotted RNA element in its 3' UTR for target-primed reverse transcription.
  • The engineered all-RNA system achieved over 80% integration efficiency in human cell lines.

Conclusions:

  • Mechanistic insights into R2 retrotransposon function were elucidated.
  • A compact, highly efficient all-RNA system for R2-mediated transgene insertion was developed.
  • This work expands the genome engineering toolbox and offers a foundation for future R2-based gene insertion tools.